Objective To investigate the autophagy capacity in clear cell renal cell carcinoma tissue compared with adjacent normal tissue.Methods Sixty-nine pairs of samples from human clear cell renal cell carcinoma tissues and relatively healthy renal tissues adjacent to the tumor were collected during surgical resection.The expressions of proteins that were participating in the regulation and execution of autophagy were detected by immunohistochemisty.Electron-microscopy was also carried out for the morphometrical analysis.Results The protein expression of p-mTOR (P =0.004),P62 (P =0.000) and ULK1 (P =0.000) were up-regulated in the carcinoma tissue,while the expression of Beclin1 (P=0.000),LC3 (P =0.000) and ATG7 (P =0.000) were down-regulated,and the expression of ATG5 (P =0.349) had no signif-icant difference compared with adjacent normal tissue.Morphometrical analysis showed that the basal autophagic activity (measured by the presence of autophagy vacuole compartment) was remarkably down-regulated in carcinoma tissue,compared with adjacent normal tissue.The expression level of mTOR was correlated with P62,LC3 and ATG7,but results showed no correlation between mTOR and Beclin 1.Conclusion Our studies show a relatively reduced autophagy capacity in clear cell renal cell carcinoma,which is regulated by multiple autophagy-related proteins such as mTOR,Beclin 1 and LC3.

Objective To observe the survival of EAE rats after recombinant adenovirus NgR specific RNA interference(Ad-shRNA-NgR) transfected the brain tissue of the experimental autoimmune encephalomyelitis(EAE),and provide the basis for EAE intervention.Methods EAE rats were randomly divided into high,medium,low and control groups(20 rats in each group).The lateral ventricle of EAE rats were injected with a titer of 1×1011 pfu/mL,1×1010 pfu/mL and 1×109 pfu/mL Ad-shRNA-NgR.The survival of EAE rats at third and seventh days after injection was observed.Results The survival rate of EAE rats of the high titer group was significantly lower than those of the middle titer group and low titer group at third and seventh days after Ad-shRNA-NgR transfected into EAE brain tissue.There was no significant difference in survival rate in middle titer group,low titer group and control group.Conclusion The titer of Ad-shRNA-NgR is safe in the experiment of EAE rats from 1×1010 pfu/mL to the range of 1×109 pfu/mL.

OBJECTIVE:To evaluate the effectiveness and safety of inhaled furosemide in the adjuvant treatment of bronchial asthma systematically,and to provide evidenced-base reference for clinical treatment. METHODS:Retrieved from PubMed,Co-chrane library,CJFD,Wanfang database and VIP,randomized controlled trials(RCTs)about routine treatment+inhaled furosemide (trial group)vs. routine treatment(control group)in the treatment of bronchial asthma were collected. After quality evaluation and data extraction according to Cochrane systematic evaluation method 5.1.0,Meta-analysis was performed with Rev Man 5.3 soft-ware. RESULTS:A total of 13 RCTs were included,involving 708 patients. The results of Meta-analysis showed that:obvious ef-fective rate [RR=1.53,95%CI(1.28,1.84),P<0.001],total effective rate [RR=1.34,95%CI(1.23,1.45),P<0.001],the inci-dence of laryngeal discomfort [RR=10.79,95%CI(1.47,78.99),P=0.02] and gastrointestinal adverse reaction [RR=10.80,95%CI (1.43,81.53),P=0.02] in trial group were all significantly higher than control group,with statistical significance. There was no sta-tistical significance in the incidence of dry mount [RR=3.71,95%CI(0.81,16.86),P=0.09] and electrolyte disorder [RR=2.38, 95%CI(0.10,56.53),P=0.59] between 2 groups. CONCLUSIONS:Based on conventional treatment,the inhaled furosemide has good therapeutic efficacy and safety for bronchial asthma,and the laryngeal discomfort and gastrointestinal reaction should be strengthened.

Objective To explore the activity and its possible neural mechanism of brain default mode network by using resting state functional magnetic resonance imaging (fMRI) in patients with mild cognitive impairment (MCI). Methods The 20 amnestic MCI patients and 25 healthy controls were included in this study, and all subjects underwent mini-mental state examination (MMSE), auditory verbal learning test (AVLT) and fMRI. The data were analyzed by amplitude of low frequency fluctuation (ALFF), and the enhanced and weakened regions of ALFF were observed and compared in both MCI patients and healthy controls. Results MMSE and AVLT tests showed that the memory function was seriously impaired in MCI patients compared with healthy controls, which is based on the short and long delayed episodic memory impairment (2.4±1.7 vs. 6.6±1.4, t=3.70, P＜0.01; 2.1±1.6 vs. 6.7±1.5, t=4.16, P＜0.01). The resting state fMRI showed that MCI patients had significant decreases of ALFF in hippocampal formation, parahippocampal cortex and lateral temporal cortex as compared with health controls (t=2.58, 2.43 and 1.75, all P＜0.01), which were closely relevant to the episodic memory. And they had significant increases in temporal-parietal joint and inferior parietal lobule (t=3.14 and 2.77, both P＜0.01). Conclusions MCI patients show significant decreased active intensity of some DMN nodes that is related to episodic memory in resting state. Increased active intensity in MCI patients would be some type of compensation.

AIM: To assess the relative agreement of GAT and NCT in IOP measurement by comparing the differences between Goldmann applanation tonometer (GAT) and non-contact tonometer (NCT) in intraocular pressure (IOP) detection.METHODS: IOP of 529 eyes of 265 volunteers were measured with both NCT and GAT, respectively.RESULTS: The measurement results of NCT were lower than that of GAT, there was significant difference between the IOP measured with NCT and GAT (19.13 vs23.43, t=22.644, P＜0.05). With the increasing of IOP values, the difference magnitude was greater, especially in IOP group that was more than 30mmHg, but the correlation coefficient became lower.CONCLUSION: The measurement results with NCT are lower than that of GAT. When the IOP with the NCT is in borderline value, it need be corrected with GAT, in order to discover the pathologically elevated IOP and avoid the misdiagnosis and mistreatment of glaucoma.

BackgroundCorneal neovascularization (CNV) is a common eye disease.The researches on the pathogenesis and treatment of CNV are focus in Ophthalmology.ObjectiveThis study was to investigate the effects of culture supernatant from human amniotic epithelial cells (AECs) on CNV in vitro and its mechanism.MethodsHuman AECs were obtained from a placenta and cultured in serum-free medium for 48 hours,and the supernatant was collected.The levels of interleukin-1 receptor antagonist (IL-1 Ra) and pigment epithelium-derived factor (PEDF) in the human AECs culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA).Rabbit corneal epithelial cells (CECs) were obtained and cultured in different concentrations of human AECs culture supernatant for 48 hours,serum-free medium was used as the control group.The expression of vascular endothelial growth factor (VEGF) mRNA and basic fibroblast growth factor (bFGF) mRNA in rabbit CECs were measured by quantitative real-time PCR (QRT-PCR).Human umbilical cord vein endothelial cells (UVECs) were cultured in the three mediums above,and the proliferation of human UVECs (absorbance value,A value) was tested by the cell counting kit 8 ( CCK8 ).Migration assay was performed by the wound healing method for the human UVECs.The membrane ultra-structure of human UVECs was examined under the atomic force microscope (AFM).ResultsCultured and passaged human AECs showed a positive response for keratin.The expression of VEGF mRNA (1.00±0.22) and bFGF mRNA (1.00±0.36) in rabbit CECs was suppressed by the human AECs culture supernatant,with a significant reduction in comparison with the serum-free DMEM group (2.98±0.46,2.55±0.48 )(P=0.001,0.002).The A value was significantly lowered in the human AECs culture group for 72 hours compared with the serum-free DMEM group ( 1.941 ± 0.036 versus 2.144 ± 0.059 ) ( P =0.000 ),and the bFGF-induced migration rate of human UVECs was strongly inhibited by the culture supernatant of human AECs in comparison with serum-free DMEM.The plasma membrane of human UVECs cultured with the human AECs culture supernatant was full of bumps,and decreased intercellular connection and cellular pseudopodia were found on the AFM image.The concentration of IL-1Ra was (153.56±0.36)ng/L and that of PEDF was (70.41 ±0.68 )μ,g/L in the human AECs culture supernatant.Nothing was deteched in serum-free DMEM group.Conclusions Human AECs culture supernatant suppressed the expression of VEGF and bFGF in CECs as well as the migration and proliferation of vascular endothelial cells.The effect may be associated with IL-1Ra and PEDF secreted by human AECs.These results suggest that human AECs may be a potential therapy for the inhibition of CNV.

Ergosterol is the important component of the fungal membrane, and having stable structure. This makes it a suitable indicator for growth of fungi. In the paper, isolation and determination techniques of ergosterol as the indicator of the fungal biomass were reviewed. The methods of extracting ergosterol include traditional saponification and refluxing, rapid physical disruption, rapid ultrasonication, supercritical fluid extraction and so on. The ergosterol determination methods are high performance liquid chromatography, gas chromatography-mass spectrometry, liquid chromatography-mass spectrometry, and thin-layer chromatography, et al. The application of these techniques was also introduced. Finally, the paper prospected the feasibility of applying the ergosterol as the indicator of fungal biomass in composting.

Armillariella tabescens EJLY2098 was induced to produce ?-mannanase with konjac fine flour (Amorphopallus rivieri) as single carbon source. This induced enzyme was then purified using DEAE ion exchange chromatography and named atMAN47. Zymologic analysis showed that the molecular weight of this ?-mannanase was approximately 47 kD. The enzyme was stable when pH ranged from 5.0 to 6.5 and could be activated by Na+ and Ba2+. With an optimal temperature of 50?C. Action mode analysis of TLC revealed that the enzyme belonged to the endo-?-mannanase family. Being a meta-acid endo-?-mannanase, it was suitable to be applied to feed industry with a promising future as an enzyme preparation.

Objective To investigate the application value of ELISA for detecting the serum anti desmoglein (Dsg) 1 and Dsg 3 in the diagnosis and treatment of pemphigus .Methods Forty‐seven patients with pemphigus in our hospital from January to De‐cember 2014 were selected as the observation group and contemporaneous 52 patients with excluding pemphigus were selected as the control group .The Dsg antibodies were detected by using indirect immunofluorescence method and Dsg 1 and Dsg3 were deter‐mined by ELISA ;their correlation with pemphigus characteristics was analyzed .Results The sensitivity and specificity of ELISA for detecting anti‐Dsg antibodies were 95 .74% and 92 .31% respectively ,while which of IIF were 93 .62% and 86 .54% respective‐ly ,showing no statistically significant difference between the two test methods (P>0 .05) .In 30 cases of pemphigus vulgaris ,16 ca‐ses (16/30) were positive Dsg1 and Dsg 3 ,8 cases of pemphigus erythematosus and 5 cases pemphigus foliaceus were positive Dsg1 only ,and 2 cases of pemphigus vegetans were both positive Dsgl and Dsg3 .The Dsgl and Dsg3 titers of pemphigus vulgaris and pemphigus vegetans were 130 .85 ± 86 and 112 .30 ± 85 .05 ,respectively ,and the disease activity score was (5 .10 ± 1 .86) points ,the correlation coefficient(r)=0 .476(P=0 .008) ,r=0 .816(P=0 .001) ,respectively .The Dsgl titer of pemphigus erythematosus and pemphigus foliaceus were 142 .59 ± 78 .52 ,and the disease activity score was (2 .77 ± 0 .92) points(r=0 .800 ,P=0 .001) .Conclu‐sion ELISA for detecting Dsg1 and Dsg3 has high sensitivity and specificity ,and is conducive to the diagnosis of pemphigus and e‐valuation of disease severity .

Traditional Chinese medicine (TCM) has definitely clinical effect in treating hyperlipidemia, but the action mechanism still need to be explored. Based on consulting Chinese Pharmacopoeia (2010), all the lipid-lowering Chinese patent medicines were analyzed by associated rules data mining method to explore high frequency herb pairs. The top three couplet medicines with high support degree were Puerariae Lobatae Radix-Crataegi Fructus, Salviae Miltiorrhizae Radix et Rhizoma-Crataegi Fructus, and Polygoni Multiflori Radix-Crataegi Fructus. The 20 main ingredients were selected from the herb pairs and docked with 3 key hyperlipidemia targets, namely 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), peroxisome proliferator activated receptor-α (PPAR-α ) and niemann-pick C1 like 1 (NPC1L1) to further discuss the molecular mechanism of the high frequency herb pairs, by using the docking program, LibDock. To construct evaluation rules for the ingredients of herb pairs, the root-mean-square deviation (RMSD) value between computed and initial complexes was first calculated to validate the fitness of LibDock models. Then, the key residues were also confirmed by analyzing the interactions of those 3 proteins and corresponding marketed drugs. The docking results showed that hyperin, puerarin, salvianolic acid A and polydatin can interact with two targets, and the other five compounds may be potent for at least one of the three targets. In this study, the multi-target effect of high frequency herb pairs for lipid-lowering was discussed on the molecular level, which can help further researching new multi-target anti-hyperlipidemia drug.
Asteraceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Humans ; Hydroxymethylglutaryl CoA Reductases ; chemistry ; genetics ; metabolism ; Hyperlipidemias ; drug therapy ; enzymology ; genetics ; metabolism ; Hypolipidemic Agents ; chemistry ; metabolism ; Membrane Proteins ; chemistry ; genetics ; metabolism ; Molecular Docking Simulation ; PPAR alpha ; chemistry ; genetics ; metabolism ; Protein Binding ; Pueraria ; chemistry